Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
acute toxicity: other routes
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1982
Report date:
1982

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
Single intraperitoneal injection with 14-day post-observation period.
GLP compliance:
no
Limit test:
yes

Test material

Constituent 1
Chemical structure
Reference substance name:
2-cyano-2-[2,3-dihydro-3-(tetrahydro-2,4,6-trioxo-5(2H)-pyrimidinylidene)-1H-isoindol-1-ylidene]-N-methylacetamide
EC Number:
278-388-8
EC Name:
2-cyano-2-[2,3-dihydro-3-(tetrahydro-2,4,6-trioxo-5(2H)-pyrimidinylidene)-1H-isoindol-1-ylidene]-N-methylacetamide
Cas Number:
76199-85-4
Molecular formula:
C16H11N5O4
IUPAC Name:
2-cyano-N-methyl-2-[3-(2,4,6-trioxotetrahydropyrimidin-5(2H)-ylidene)-2,3-dihydro-1H-isoindol-1-ylidene]acetamide
Details on test material:
- Name of test material (as cited in study report): Pigmentgelb HNN 79/195
- Physical state: solid, yellow
- Analytical purity: 88 %
- Impurities (identity and concentrations): 10 % 2-cyano-2-[2,3-dihydro-3-oxo-1H-isoindol-1-ylidene]-N-methylacetamide
- Stability: At least 2 years under normal conditions

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Dr. K. Thomae GMBH, Biberach, Germany
- Age at study initiation: 12 weeks
- Mean weight at study initiation: 170 g (males), 180 g (females)
- Fasting period before study: 16 hours before administration, but water is available ad libitum.
- Housing: 5 per cage, Stainless steel wire mesh cages, type DK-III
- Diet (e.g. ad libitum): Ssniff R; FA. SSNIFF, Soest, Germany
- Water: Demineralized water each workday, tap water on holidays; ad libitum
- Acclimation period: at least 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-26
- Humidity (%): 45-75
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
intraperitoneal
Vehicle:
CMC (carboxymethyl cellulose)
Remarks:
0.5% aqueous CMC-solution
Details on exposure:
VEHICLE
- Concentration in vehicle: 20 % in 0.5% aqueous carboxymethyl cellulose

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg
Doses:
2000 mg/kg
No. of animals per sex per dose:
5
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: weighing: before administration, then on days 3, 7 and 13;
cageside observations: < 15', 15', 30', 1 h, 2 h, 4 h and 5 h after administration of the test substance and then once each workday.
Check for moribund and dead animals twice each workday and once on holidays.
- Necropsy of survivors performed: yes, withdrawal of food 16 houre before sacrifice; necropsy with gross-pathological examination.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Remarks on result:
other: No mortality.
Mortality:
No mortality observed.
Clinical signs:
Males and females: Dyspnea, apathy, staggering, tonus with stretching, urine yellow/orange, poor general state
Females: Additionally, piloerection
Body weight:
The body weight of the animals showed no adverse effects in comparison with that of the control.
Mean body weight (male / female)
- Before the study
Treatment group: 185 / 170
- After 3 days
Treatment group: 194 / 180
- After 7 days
Treatment group: 213 / 216
- After 13 days
Treatment group: 225 / 264
Control: 367 / 249
Gross pathology:
Sacrificed animais (male and female): abdominal organs, particularly Glisson's capsule, severely stained by the test substance; focal adhesions of the lobes of the liver to each other, in some cases with rounding of the margins of the liver.
Other findings:
No other findings.

Applicant's summary and conclusion

Conclusions:
In conclusion, the median lethal dose (LD50) after intraperitoneal injection in rats was determined to be > 2000 mg/kg bw. 
Executive summary:

In this non-guideline study, 5 rats per sex were administered a single dose (2000 mg/kg bw in 0.5% aqueous CMC-solution) of the test substance via intraperitoneal injection. Clinical signs were monitored and necropsy was carried out at termination or after death if an animal died during the study. The observation period was until day 14 after application.


No mortality was observed.


Clinical signs were dyspnea, apathy, staggering, tonus with stretching, yellow/orange colored urine, poor general state in males and femals. Additionally, females also showed piloerection. The body weight of the animals showed no adverse effects in comparison with that of the control. After sacrifice, abdominal organs, particularly Glisson's capsule, were severely stained by the test substance in both males and females. No other findings were recorded. 


In conclusion, the median lethal dose (LD50) after intraperitoneal injection in rats was determined to be > 2000 mg/kg bw.